Inhibitors of inflammation and reperfusion injury and methods of use thereof

ABSTRACT

The invention provides a novel class of substituted isoindolinone derivatives. Pharmaceutical compositions, and methods of making and using the compounds, or pharmaceutically acceptable salts, hydrates, prodrugs, or mixtures thereof are also described.

RELATED APPLICATIONS

[0001] This application claims priority to U.S. Ser. No. 60/195,622,filed Apr. 6, 2000. The contents of this application are incorporated byreference in their entirety.

STATEMENT OF GOVERNMENT SUPPORT

[0002] This invention was made with United States Government supportunder grant number R44NS37642-02, awarded by the National Institute ofNeurological Disorders and Stroke (NINDS). The United States Governmenthas certain rights in the invention.

FIELD OF THE INVENTION

[0003] The invention relates generally to inhibitors of inflammation andreperfusion injury. In particular, the invention relates to2,3-dihydro-isoindol-1-one derivatives and nucleoside analogs, and moreparticularly to nucleoside-isoindolinone conjugates.

BACKGROUND OF THE INVENTION

[0004] Inflammation disorders, such as arthritis, colitis, andautoimmune diabetes typically manifest themselves as disorders distinctform those associated with reperfusion injury, e.g., stroke and heartattack, and can present clinically as different entities. However, therecan be common underlying mechanisms between these two types ofdisorders. In particular, inflammation and reperfusion injury can induceproinflammatory cytokine and chemokine synthesis. Induction ofpro-inflammatory cytokines can, in turn, result in production ofcytotoxic free radicals such as nitric oxide (NO) and superoxide. Nitricoxide and superoxide can react to form peroxynitrite (ONOO⁻). Szabó etal, Shock 6:79-88, 1996.

[0005] Peroxynitrite-induced cell necrosis observed in inflammation andreperfusion injury involves, in significant part, the activation of thenuclear enzyme poly (ADP-ribose) synthetase (PARS). Activation of PARSis thought to be an important step in the cell-mediated death observedin inflammation and reperfusion injury. Szabó et al., Trends Pharmacol.Sci. 19: 287-98, 1998.

[0006] A number of PARS inhibitors have been described in the art. See,e.g., Banasik et al., J. Biol. Chem., 267:1569-75, 1992, and Banasik etal., Mol. Cell. Biochem., 138:185-97, 1994. Additionally, some potentPARS inhibitors are reported in, for example, WO 00/39104, WO 00/39070,WO 99/59975, WO 99/5973, WO 99/11649, WO 99/11645, WO 99/11644, WO99/11628, WO 99/11628, WO 99/11623, WO 99/11311, WO 00/42040; Zhang etal., Biochem. Biophys. Res. Commun., 278:590-98, 2000, White et al.,J.Med. Chem., 43:4084-4097, 2000; Griffin et al., J. Med. Chem.,41:5247-5256, 1998; Shinkwin et al., Bioorg. Med. Chem., 7:297-308,1999. Furthermore, side effects of some of the best known-PARPinhibitors have been discussed in Milan et al, Science, 223:589-591,1984.

[0007] Certain isolindolinone derivatives are known in the art. Forexample, inhibitors of platelet aggregation are reported in Egbertson etal., J. Med. Chem., 42:2409-21, 1999; dopamine D4 receptorisoindolinones are reported in Belliotti et al., Bioorg. Med. Chem.Lett., 8:1499-502, 1998; antipsychotic agents are disclosed in Norman etal., J. Med. Chem., 37: 2552-63, 1994 and in Normal et al., J. Med.Chem., 36: 3417-23, 1993. The antiarrhythmic activity of isoindolione isshown in Dugger et al. Drug Metab. Dispos. 4:262-268, 1976, andsubstituted 2,3-dihydro-1H-isoindol-1-one derivatives for treatinghyperlipemia remedy are disclosed in WO 98/54135.

[0008] Syntheses of substituted 2,3-dihydroisoindolinones, other thanthe compounds of the invention, are reported in, for example, Duckworthet al., J. Chem. Soc., Perkin Trans. 1:815-21, 1996; Kamochi et al.,Daiichi Yakka Daigaku Kenkyu Nenpo 20:1-10, 1989; McAlees et al., J.Chem. Soc. Perkin Trans 1, 1:2038-2040, 1977; Tomita et al., J. Chem.Soc. C, 2:183-8, 1969; Do Minh et al., J. Org. Chem., 42:4217-4221,1977; and O'Sullivan et al., J. Chem. Soc. Chem. Commun., 17:1165-1166,1984.

[0009] Various nucleoside peptides and amide derivatives are shown in,for example, Kawana et al. , J. Org. Chem., 37:288-91 (1972); U.S. Pat.Nos. 3,864,483; 3,914,414; 3,914,415; 3,966,917; 4,029,884; and inGerman patents DE 2417465 and DE 2213180.

SUMMARY OF THE INVENTION

[0010] The invention is based in part on the discovery of novelcompounds and their unexpected effects in inhibiting inflammation and intreating reperfusion injuries.

[0011] Accordingly, one aspect of the invention includes novelsubstituted isoindolinone derivatives. In another aspect, the inventionrelates to substituted nucleoside analogs. In yet another aspect, theinvention includes a conjugate according to Formula I, as set forth inthe Detailed Description of the Invention, below.

[0012] Also provided by the invention is a method of treatinginflammatory and reperfusion conditions in mammals by administering to amammal in need of such treatment an effective amount of the compounds ofthe invention, for example, a conjugate according to Formula I.

[0013] In a further aspect, the invention also includes a method for theproduction of the compounds of the invention.

[0014] In one aspect of the invention, a nucleoside or nucleoside analogis conjugated to a compound that is useful for inhibiting inflammationor for treating reperfusion injuries. In some embodiments, thenucleoside moiety increases the anti-inflammatory or anti-reperfusionactivity of the conjugated compound. In a particular embodiment, anucleoside moiety is conjugated to an isoindolinone moiety.

[0015] The compounds described in the current invention are potentcompounds that can be used to treat a variety of conditions anddiseases, typically those known to involve inflammatory mediatorproduction and cell death.

[0016] The details of one or more embodiments of the invention are setforth in the accompanying description below. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. Other features, objects, and advantagesof the invention will be apparent from the description and from theclaims. In the specification and the appended claims, the singular formsalso include the plural unless the context clearly dictates otherwise.Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. All patents and publicationscited in this specification are incorporated by reference.

DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention provides a novel class of substitutednucleoside derivatives according to Formula I:

A—Z₁—L—Z₂—G  I

[0018] or a pharmaceutically acceptable prodrug, hydrated salt, ormixtures thereof, wherein A and G are connected via Z₁ and Z₂,

[0019] Wherein

[0020] X₁ and X₂ are, independently, N or CH;

[0021] X₃ is CR₇═CR₈, CHR₇—CHR₈, CR₇═N, N═CR₇, N═N, NR₇—O, CHR₇—O orCHR₇—S, where R₇ and R₈ are, independently, H, alkyl, amino, hydroxy,alkoxy;

[0022] R₁ and R₂ are, independently, H, alkyl, NH₂, OH, SH, Cl, NHR₉,N═R₉, N═NR₉, or amide, where R₉ is alkyl, aryl, arylalkyl,alkyl-heterocycle;

[0023] R₃ and R₄ are independently H, C1-5 alkyl, hydroxy, amino orhalo;

[0024] R₅ and R₆ are, independently, alkyl, other acyl, or R₅ and R₆,taken together, form a 5-or 6-membered, substituted or unsubstitutedheterocycle.

[0025] In some embodiments, A is a 5′ modified purine or pyrimidinenucleoside, or a derivative thereof (e.g., where X₁═X₂═N, X₃ is N═CH, R₁and R₂ are NH₂ or OH).

[0026] Z₁ is —CH₂O—, —CH₂NR₁₀—, —CH₂NR₁₀C(O)—, —CONR₁₀—, —CO₂—,—CH₂NHCONH—, —CH₂—, —CH₂NHCSNH—, —CO—, —CH₂CO₂—, —NHCO₂, S, SO₂, CH₂S,SO;

[0027] Z₂ is; —NR₁₀CO—, —C(O)NR₁₀—, —NHCONH—, —OC(O)—, —C(O)O—, NHCS,—CSNH—, NHCSNH, O, CO, OCO, OCONH, NH, CH₂, CH-alkyl, NHCO₂, S, SO₂, CS,SO.

[0028] It is understood that tautomeric forms, where possible, areincluded in the invention, and that where tautomerisation is possible,the tautomer represented herein as structures II or III may notrepresent the dominant tautomer.

[0029] The linker, L, can be H, O, S, C1-15 alkylene chain, which can besubstituted in one or more positions, or a 5, 6 or 7-membered carbocycleor heterocycle (optionally substituted in one or more positions),provided that when Z₁ is O, L is not H, and when L is H, Z₂ and G areabsent. In some embodiments, L is substituted with amino, alkyl, halo,hydroxy, thio, or epoxide groups in any combination;

[0030] In some embodiments, L may contain:

[0031] i) one or more heteroatoms chosen from N, O, S, alone or in anyacceptable combination, including, but not limited to, SO₂, S—S, N═N;

[0032] ii) 5 or 6 member cyclic moieties, for example, hetercyclic,carbocyclic, aromatic or otherwise, with no restriction with respect topoints of attachment to L, or to Z₁ or Z₂, where applicable. The cyclicmoieties can be either unsubstituted or substituted with lower alkyl,hydroxy, keto, amino, aminoalkyl, halo, alkoxy groups. Examples ofacceptable cycles include substituted aryl, substituted heterocyclic orheterocyclic amines such as piperidine, piperazine, pyrole, imidazole,benzimidazole, tetrazoles, indole, isoquinoline, quinoline, pyrrolidine;

[0033] iii) varying degrees of unsaturation, including alkene, imine,diazo; or

[0034] iv) combinations of i, ii, and iii.

[0035] In some embodiments, G can be H, OH, SH, NH₂, CO₂H, or asubstituted alkyl, aryl, alkylaryl, carbocyclic, heterocyclic, bicyclic,a bicyclic heterocycle, biphenyl or heterocyclic amine such as, forexample, a substituted or unsubstituted piperidine, piperazine, pyrole,imidazole, benzimidazole, tetrazole, indole, isoquinoline, quinoline,pyrrolidine.

[0036] In some aspects of the invention, G itself is an inhibitor ofinflammation or of reperfusion injury. For example, G can be anyinhibitor of PARS whose potency as an inhibitor of PARS is preferablyincreased by incorporation into a structure as indicated by Formula I.It will be recognized to those skilled in the art that the site ofattachment of the linker, L, to moiety G (via Z₂) and the nature of Land Z₂, will influence the overall potency of the conjugate as aninhibitor of PARS.

[0037] In some aspects of the invention, moiety G can be a member of anovel class of isoindolone compounds represented by Formula IV,described below.

[0038] One aspect of the invention includes a compound according toFormula IV, unconjugated to the A moiety.

[0039] Specifically, the present invention relates to a compound ofFormula IV, wherein:

[0040] Y is O, S, Se, NH, N-alkyl, or N-aryl;

[0041] R₁₁ is H, OH, aryl, alkyl, or an amino acid side chain;

[0042] R₁₂ and R₁₃ are, independently, a hydrogen, alkyl, aryl,heterocycle, OH, O-alkyl, O-aryl, N-alkyl, N-aryl, or, taken together,are ═O, ═NH, ═S; and

[0043] R₁₄, R₁₅, R₁₆, and R₁₇ are, independently, hydrogen, halo,alkylhalo, hydroxy, alkoxy, C1-C10 straight or branched chain alkyl,C2-C10 straight or branched chain alkenyl group, C3-C8 carbocyclic,aryl, alkylamino, amino, carboxy, ester, arylalkyl, or nitro.

[0044] In certain embodiments, R₁₄, R₁₅, R₁₆, and R₁₇ are,independently, H, Z₂—L—Z₁-A (as defined above), or Q—B—D, wherein:

[0045] Q is NHCO, NHCONH, O, CO, OCO₂, OCO, OCONH, NH, CH₂, CH-alkyl,NHCO₂, S, SO₂, CS, or SO;

[0046] B is C1-C10 straight or branched chain alkyl, C2-C 10 straight orbranched chain alkenyl group, C3-C8 carbocyclic, aryl, alkylamino,amino, alkylamido, arylamido, carboxy, ester, anhydride, or an arylalkylgroup substituted with one or more hydrogen, halogen, alkylhalo,hydroxy, nitro, amino, amido, carbamate, or carbonate groups; and

[0047] D is hydrogen, a substituted heterocycle or carbocycle, or astraight or branched chain alkyl amine. In some embodiments, thesubstituents can be: hydrogen, alkylhalo, alkylhydroxy, C1-C10 straightor branched chain alkyl, C2-C10 straight or branched chain alkenyl,C2-C10 straight or branched chain alkynyl, C3-C8 carbocyclic, aryl,benzyl, alkylamino, alkylamido, alkylcarboxy, alkylester, arylalkyl, orcyclic heterocyclic amines. Substituted amines cyclic or heterocyclicamines include piperidine, piperazine, N-alkylated or alkylcarbonylatedpiperazines, pyrole, imidazole, benzimidazole, tetrazoles, indole,isoquinoline, quinoline, pyrrolidine, aniline, substituted aniline,purine, nucleosides, nucleotides, sugars, hydroxylated alkanes,glycerol, and other C2 to C10 branched or cyclic or cycloalkenyl aminesor heterocyclic compounds.

[0048] In some embodiments, D is absent.

[0049] Also included in the invention are compounds according to FormulaIV, wherein R₁₃ and R₁₄ form a heterocyclic or a carbocyclic ringcontaining 5 to 10 members, e.g., 5, 6, 7, 8, 9, or 10 members.Alternatively, R₁₄ and R₁₅ can be joined to form a 5 to 10 memberheterocyclic or carbocyclic ring.

[0050] The invention also includes a pharmaceutical composition thatincludes a compound of the invention and a pharmaceutically acceptablecarrier. For example, the invention includes a compound according toFormula I provided as a pharmaceutically acceptable prodrug, hydratedsalt, or mixtures thereof.

[0051] Salts encompassed within the term “pharmaceutically acceptablesalts” refer to non-toxic salts of the compounds of this invention whichare generally prepared by reacting the free base with a suitable organicor inorganic acid to produce “pharmaceutically acceptable acid additionsalts” of the compounds described herein. These compounds retain thebiological effectiveness and properties of the free bases.Representative salts include, e.g., water-soluble and water-insolublesalts, such as the acetate, amsonate(4,4-diaminostilbene-2,2′-disulfonate), benzenesulfonate, benzonate,bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calciumedetate, camsylate, carbonate, chloride, citrate, clavulariate,dihydrochloride, edetate, edisylate, estolate, esylate, fumarate,gluceptate, gluconate, glutamate, glycollylarsanilate,hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate,lactobionate, laurate, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate,N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate,oxalate, palmitate, pamoate (1,1-methylene-bis-2-hydroxy-3-naphthoate,embonate), pantothenate, phosphate/diphosphate, picrate,polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate,subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate,tartrate, teoclate, tosylate, triethiodide, and valerate salts.

[0052] Methods of Using Substituted Nucleoside Derivatives

[0053] The invention also includes a method of inhibitingpoly(ADP)-ribose synthase activity (PARS) in a cell. This enzyme, whichis also known as poly(ADP-ribose)synthetase and PARP (poly(ADP-ribose)polymerase, EC 2.4.99), and ADP-ribosyltransferase (ADPRT, EC 2.4.2.30),is a nuclear enzyme that catalyzes a transfer of the ADP ribose moietyof NAD+ to an acceptor protein.

[0054] The method includes contacting the cell with a compound ofFormula I or IV in an amount sufficient to inhibit poly(ADP)-ribose-synthase in the cell. In general, any cell having, orcapable of having, PARS activity, can be used. The cell can be providedin any form as long as it is accessible to the compound. For example,the cell can be provided in vitro, ex vivo, or in vivo. PARS activitycan be measured using any method known in the art, e.g., methods asdescribed in Banasik et al, J. Biol. Chem. 267:1569-75 (1991).

[0055] Also provided in the invention is a method of inhibiting,preventing, or treating inflammation in a subject. The inflammation canbe associated, e.g., with an inflammatory disease. Inflammatory diseasesrefer to diseases or conditions where there is an inflammation of thebody tissue. These include local inflammatory responses and systemicinflammation. Examples of such diseases and conditions include:transplant rejection; chronic inflammatory disorders of the joints,including arthritis, rheumatoid arthritis, osteoarthritis and bonediseases associated with increased bone resorption; inflammatory boweldiseases such as ileitis, ulcerative colitis, Barrett's syndrome, andCrohn's disease; inflammatory lung disorders such as asthma, adultrespiratory distress syndrome, and chronic obstructive airway disease;inflammatory disorders of the eye including corneal dystrophy, trachoma,onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis;chronic inflammatory disorders of the gum, including gingivitis andperiodontitis; tuberculosis; leprosy; inflammatory diseases of thekidney including uremic complications, glomerulonephritis and nephrosis;inflammatory disorders of the skin including sclerodermatitis, psoriasisand eczema; inflammatory diseases of the central nervous system,including chronic demyelinating diseases of the nervous system, multiplesclerosis, AIDS-related neurodegeneration and Alzheimer's disease,infectious meningitis, encephalomyelitis, Parkinson's disease,Huntington's disease, amyotrophic lateral sclerosis and viral orautoimmune encephalitis; autoimmune diseases including diabetesmellitus, immune-complex vasculitis, systemic lupus erythematosus (SLE);and inflammatory diseases of the heart such as cardiomyopathy, ischemicheart disease hypercholesterolemia, atherosclerosis; as well as variousother diseases with significant inflammatory components, includingpreeclampsia; chronic liver failure; brain and spinal cord trauma, andcancer. There may also be a systemic inflammation of the body,exemplified by gram-positive or gram negative shock, hemorrhagic oranaphylactic shock, or shock induced by cancer chemotherapy, e.g., shockassociated with pro-inflammatory cytokines.

[0056] The invention also includes a method of treating, preventing, orotherwise inhibiting reperfusion injury in a subject in need oftreatment, prevention, or inhibition thereof. The method includesadministering a compound of the invention in an amount sufficient toinhibit reperfusion injury in the subject. Reperfusion refers to theprocess whereby blood flow in the blood vessels is resumed after bloodflow has been interrupted, such as occurs following constriction orobstruction of the vessel. Reperfusion is typically associated withischemia and may result following a naturally occurring episode, such asa myocardial infarction or stroke, or during a surgical procedure whereblood flow in vessels is purposely or unintentionally blocked off.

[0057] The subject in the above-mentioned methods can be, e.g., amammal, e.g., a human, mouse, rat, dog, cat, horse, cow, pig, ornon-human primate. Administration can be systemic or topical, and can beprophylactic or therapeutic.

[0058] The term “pharmacologically effective amount” means that amountof a drug or pharmaceutical agent that will elicit the biological ormedical response of a tissue, system, animal or human that is beingsought by a researcher or clinician.

[0059] The invention also includes pharmaceutical compositions suitablefor inhibiting or preventing inflammation or reperfusion injury, PARSactivity, or more than one of these activities. In practice, thecompounds or their pharmaceutically acceptable salts, are administeredin amounts which will be sufficient to inhibit inflammatory conditionsor disease and/or prevent the development of inflammation orinflammatory disease in a subject, such as a mammal, and are used in theform most suitable for such purposes. The compositions are preferablysuitable for internal use and include an effective amount of apharmacologically active compound of the invention, alone or incombination, with one or more pharmaceutically acceptable carriers. Thecompounds are especially useful in that they have very low, if anytoxicity.

[0060] The compounds herein described can form the active ingredient ofa pharmaceutical composition, and are typically administered inadmixture with suitable pharmaceutical diluents, excipients or carriers(collectively referred to herein as “carrier” materials) suitablyselected with respect to the intended form of administration, that is,oral tablets, capsules, elixirs, syrups and the like. The compositionstypically will include an effective amount of active compound or thepharmaceutically acceptable salt thereof, and in addition, and may alsoinclude any carrier materials as are customarily used in thepharmaceutical sciences. Depending on the intended mode ofadministration, the compositions may be in solid, semi-solid or liquiddosage form, such as, for example, injectables, tablets, suppositories,pills, time-release capsules, powders, liquids, suspensions, or thelike, preferably in unit dosages.

[0061] Administration of the active compounds and salts described hereincan be via any of the accepted modes of administration for therapeuticagents. These methods include systemic or local administration such asoral, nasal, parenteral, transdermal, subcutaneous, or topicaladministration modes.

[0062] For instance, for oral administration in the form of a tablet orcapsule (e.g., a gelatin capsule), the active drug component can becombined with an oral, non-toxic pharmaceutically acceptable inertcarrier such as ethanol, glycerol, water, and the like. Moreover, whendesired or necessary, suitable binders, lubricants, disintegratingagents and coloring agents can also be incorporated into the mixture.Suitable binders include starch, magnesium aluminum silicate, starchpaste, gelatin, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone, natural sugars such as glucose or beta-lactose,corn sweeteners, natural and synthetic gums such as acacia, tragacanthor sodium alginate, polyethylene glycol, waxes and the like. Lubricantsused in these dosage forms include sodium oleate, sodium stearate,magnesium stearate, sodium benzoate, sodium acetate, sodium chloride,silica, talcum, stearic acid, its magnesium or calcium salt and/orpolyethyleneglycol and the like. Disintegrators include, withoutlimitation, starch, methyl cellulose, agar, bentonite, xanthan gumstarches, agar, alginic acid or its sodium salt, or effervescentmixtures, and the like. Diluents, include, e.g., lactose, dextrose,sucrose, mannitol, sorbitol, cellulose and/or glycine.

[0063] The compounds of the invention can also be administered in suchoral dosage forms as timed release and sustained release tablets orcapsules, pills, powders, granules, elixers, tinctures, suspensions,syrups and emulsions.

[0064] Liquid, particularly injectable compositions can, for example, beprepared by dissolving, dispersing, etc. The active compound isdissolved in or mixed with a pharmaceutically pure solvent such as, forexample, water, saline, aqueous dextrose, glycerol, ethanol, and thelike, to thereby form the injectable solution or suspension.Additionally, solid forms suitable for dissolving in liquid prior toinjection can be formulated. Injectable compositions are preferablyaqueous isotonic solutions or suspensions. The compositions may besterilized and/or contain adjuvants, such as preserving, stabilizing,wetting or emulsifying agents, solution promoters, salts for regulatingthe osmotic pressure and/or buffers. In addition, they may also containother therapeutically valuable substances.

[0065] The compounds of the present invention can be administered inintravenous (both bolus and infusion), intraperitoneal, subcutaneous orintramuscular form, all using forms well known to those of ordinaryskill in the pharmaceutical arts. Injectables can be prepared inconventional forms, either as liquid solutions or suspensions.

[0066] Parental injectable administration is generally used forsubcutaneous, intramuscular or intravenous injections and infusions.Additionally, one approach for parenteral administration employs theimplantation of a slow-release or sustained-released systems, whichassures that a constant level of dosage is maintained, according to U.S.Pat. No. 3,710,795, incorporated herein by reference.

[0067] Furthermore, preferred compounds for the present invention can beadministered in intranasal form via topical use of suitable intranasalvehicles, or via transdermal routes, using those forms of transdermalskin patches well known to those of ordinary skill in that art. To beadministered in the form of a transdermal delivery system, the dosageadministration will, of course, be continuous rather than intermittentthroughout the dosage regimen. Other preferred topical preparationsinclude creams, ointments, lotions, aerosol sprays and gels, wherein theconcentration of active ingredient would range from 0.1% to 15%, w/w orw/v.

[0068] For solid compositions, excipients include pharmaceutical gradesof mannitol, lactose, starch, magnesium stearate, sodium saccharin,talcum, cellulose, glucose, sucrose, magnesium carbonate, and the likemay be used. The active compound defined above, may be also formulatedas suppositories using for example, polyalkylene glycols, for example,propylene glycol, as the carrier. In some embodiments, suppositories areadvantageously prepared from fatty emulsions or suspensions.

[0069] The compounds of the present invention can also be administeredin the form of liposome delivery systems, such as small unilamellarvesicles, large unilamellar vesicles and multilamellar vesicles.Liposomes can be formed from a variety of phospholipids, containingcholesterol, stearylamine or phosphatidylcholines. In some embodiments,a film of lipid components is hydrated with an aqueous solution of drugto a form lipid layer encapsulating the drug, as described in U.S. Pat.No. 5,262,564.

[0070] Compounds of the present invention may also be delivered by theuse of monoclonal antibodies as individual carriers to which thecompound molecules are coupled. The compounds of the present inventionmay also be coupled with soluble polymers as targetable drug carriers.Such polymers can include polyvinylpyrrolidone, pyran copolymer,polyhydroxypropyl-methacrylamide-phenol,polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysinesubstituted with palmitoyl residues. Furthermore, the compounds of thepresent invention may be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example,polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross-linked or amphipathic block copolymers of hydrogels.

[0071] If desired, the pharmaceutical composition to be administered mayalso contain minor amounts of non-toxic auxiliary substances such aswetting or emulsifying agents, pH buffering agents, and other substancessuch as for example, sodium acetate, triethanolamine oleate, etc.

[0072] The dosage regimen utilizing the compounds is selected inaccordance with a variety of factors including type, species, age,weight, sex and medical condition of the patient; the severity of thecondition to be treated; the route of administration; the renal andhepatic function of the patient; and the particular compound or saltthereof employed. An ordinarily skilled physician or veterinarian canreadily determine and prescribe the effective amount of the drugrequired to prevent, counter or arrest the progress of the condition.

[0073] Oral dosages of the present invention, when used for theindicated effects, will range between about 0.05 to 1000 mg/day orally.The compositions are preferably provided in the form of scored tabletscontaining 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100.0, 250.0,500.0 and 1000.0 mg of active ingredient. Effective plasma levels of thecompounds of the present invention range from 0.002 mg to 50 mg per kgof body weight per day.

[0074] Compounds of the present invention may be administered in asingle daily dose, or the total daily dosage may be administered individed doses of two, three or four times daily.

[0075] Any of the above pharmaceutical compositions may contain 0.1-99%,1-70%, or, preferably, 1-50% of the active compounds of the invention asactive ingredients.

[0076] Methods of Making the Compounds of the Invention

[0077] Examples of synthetic pathways for making compounds according tothe invention are set forth in the Examples below. For example toprepare isoindolinone compounds according to Formula IV, 3-nitro,3-fluoro, 3-hydroxy or 3-ethylcarbonate substituted esters are preparedby standard esterification methods (e.g., SOCl₂/alcohol andalcohol/acid) and treated with NBS to make corresponding2-bromomethylene benzoate derivatives. The 4-substituted isoindolinonesare prepared by reacting a methanolic solution of ammonia with acorresponding 2-bromomethylene benzoic acid esters. The4-nitroisoindolinones are reduced to 4-amino isoindolinones by ahydrogenation reaction, and then treated with various acid chlorides,anhydrides or isocyanates to generate amide and carbamate derivatives.The 4-N-chloroacetyl and other C-4 substituted derivatives can betreated with various amines, acid chlorides or other electrophiles toyield compounds according to Formula IV.

[0078] The 4-aminoisoindolinone acid derivatives (compounds 54-64) aretreated with 2′,3′-isopropylidene-5′-aminomethyl adenosine in presenceof 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) togenerate adenosine amide derivatives, and then the protected adenosineamides are deprotected using TFA and water to yield compounds accordingto Formula IV (compounds 101-108 and 111-120). Lee et al., Bioorg. &Med. Chem. Lett., 9:1365-1370, 1999. The 4-amino derivatives (compounds65-74) are treated with 2′,3′-isopropylidene-adenosine-5′-carboxylicacid in presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (EDCI) or dicyclohexylcarbodiimide (DCC) to generate2′,3′-isopropylideneadenosine amide derivatives, and then theisopropylidene group is deprotected using TFA and water to yieldcompounds according to Formula IV (compounds 123-130) and 111-120).Kawana et al., J. Org. Chem. 37:288-291, 1972.

[0079] It is to be understood that while the invention has beendescribed in conjunction with the detailed description thereof, theforegoing description is intended to illustrate and not limit the scopeof the invention. The invention will be further described in thefollowing examples, which do not limit the scope of the inventiondescribed in the claims. The following examples illustrate the synthesisof novel compounds of the invention, and of the use of these compoundsto inhibit inflammation and reperfusion.

EXAMPLES Example 1

[0080] Synthesis of Substituted 2,3-dihydro-isoindol-1-ones

[0081] a) General Methods

[0082]¹NMR spectra were obtained from Varian 300 MHz spectrophotometerand chemical shift is reported in parts per million (ppm, δ). TLC wascarried out on precoated TLC plates with silica gel 60 F-254 andpreparative TLC on precoated Whatman 60A TLC plates. All intermediatesand final compounds were characterized on the basis of ¹NMR and massspectrometry (MS) data. An exemplary synthesis of anucleoside-isoindolinone conjugate is outlined in Scheme 1.

[0083] b) Synthesis of methyl-2-bromomethyl-3-nitro-benzoate:

[0084] To a well stirred solution of methyl-4-nitro-2-methyl-benzoate (5gm, 0.025 mol.) in carbon tetrachloride or CH₂Cl₂ (75 ml) was addedN-bromo-succinimide (NBS) (4.950 gm, 0.03 mol) and AIBN (50 mg) and thehomogeneous reaction mixture was refluxed for 16 hr or until thestarting material had completely reacted. The reaction mixture was thencooled to room temperature, and the succinimide was removed byfiltration. The filtrate was concentrated to give crude product, whichafter treatment with hexane providedmethyl-2-bromomethyl-3-nitro-benzoate as a white solid (6.9 gm, 98%yield).

[0085] c) Synthesis of 4-nitro-2,3-dihydro-1H-isoindol-1-one (3):

[0086] To a solution of methyl-2-bromomethyl-3-nitro-benzoate (6.9 gm,0.025 mol.) in methanol, was added slowly a methanolic solution ofammonia (50 ml), and the reaction mixture was stirred at roomtemperature for 2 hr. The pale yellow colored solid separated out wasfiltered, dried under vacuum to give4-nitro-2,3-dihydro-1H-isoindol-1-one (compound 3, 3.570 gm, 81%).

[0087]¹HNMR (DMSO-D₆): 4.73 (s, 2H), 7.66 (dd, 1H), 8.03 (d, 1H), 8.32(d, 1H), 8.8 (bs, 1H).

[0088] d) Synthesis of 4-amino-2,3-dihydro-1H-isoindol-1-one (4):

[0089] To a solution of 4-nitro-2,3-dihydro-1H-isoindol-1-one (compound3, 3.2 gm, 0.018 mmol.) in DMF (10 ml), was added ammonium formate(5.750 gm), and Pd—C (100 mg), and the reaction mixture was stirred at100° C. for 30 min. The mixture was then filtered through a pad ofcelite, and the celite was washed with DMF (10 ml) and water (10 ml).The filtrate was concentrated under vacuum to give compound 4 (2.2 gm,83%).

[0090]¹NMR (DMSO-D₆): 4.20 (s, 2H, CH₂), 5.20 (bs, NH₂), 6.82 (dd,1H,Ar—H), 7.02 (dd,1H, Ar—H), 7.16 (t,1H, Ar—H), 8.20 (bs,1H).

[0091] e) General Synthesis of 2,3-dihydro-1H-isoindol-1-ones (5, 76,and 77) from compound 3:

[0092] To a solution compound 3 (3.250 gm, 0.018 mmol.) in DMF (20 ml),was added ammonium formate (5.750 gm), and Pd—C (100 mg), and thereaction mixture was stirred at 100° C. for 30 min. The mixture was thenfiltered through a pad of celite, and the celite was washed with DMF (10ml) and water (10 ml). The filtrate was transferred into a 250 ml roundbottom flask along with 10 ml ethyl acetate, and treated with an aqueousNaHCO₃ solution, followed by the addition of excess chloroacetylchloride at room temperature. The reaction mixture was stirred at roomtemperature (RT) for 30 min. A solution of saturated KHCO₃ was thenadded and the resultant solid was filtered and dried under vacuum togive 3-N-chloroacetyl-2,3-dihydro-1H-isoindol-1-one, 5 (3.2 gm, 80%).Using similar methods, chloropropanyl, and chlorobutanyl derivativeswere synthesized.

[0093] f) Synthesis of4-N,N-dialkylacetamido-2,3-dihydro-1H-isoindol-1-ones (6-8,11-24, and68-72):

[0094] To a solution of 4-chloroacetyl-2,3-dihydro-1H-isoindol-1-one 6(0.5 gm) in DMF or CH₃OH (5 ml) a solution of amine in methanol or DMF(5 ml) was added, and the reaction mixture was stirred at roomtemperature for 2-24 hr. The reaction mixture was dried under vacuum andthe resultant solid was filtered, and then recrystalized fromCH₃OH-ether to give 4-aminocompounds. The HCl salt of isoindolinone 7was prepared using a solution of HCl in ether (2 M, 10 ml) and CH₃OH.The resulting solid was filtered and washed thoroughly with dry ether,and recrystalized from methanol-ether.

[0095] g) Synthesis of4-N-(N-methylpiperazine)propylamido-2,3-dihydro-1H-isoindol-1-one (9):

[0096] To a solution of 4-chloropropanyl-2,3-dihydro-1H-isoindol-1-one(compound 76, 0.140 gm, 0.58 mmol) in CH₃OH (1 ml) a solution ofN-methyl piperazine (5 ml) was added and the reaction mixture wasstirred at room temperature for 1 hr. A solid formed and was separatedby filtration, then washed thoroughly with dry ether, and recrystalizedfrom methanol-ether to provide compound 9 (0.110 gm, 62%).

[0097] h) Synthesis of4-N-(N-methylpiperazine)butylamido-2,3-dihydro-1H-isoindol-1-one (10):

[0098] To a solution of 4-chlorobutyryl-2,3-dihydro-1H-isoindol-1-one(Compound 77, 0.100 gm, 0.39 mmol) in CH₃OH (1 ml) a solution ofN-methyl piperazine (5 ml) was added and the reaction mixture wasstirred at room temperature for 1 hr. The precipitate was filtered andwashed thoroughly with dry ether, and recrystalized from methanol-etherto provide 10 (0.045 gm, 36%).

[0099] i) Synthesis of 3-methylcarbonate-2-bromomethyl-methylbenzoate:

[0100] To a solution of 3-hydroxy-2-methyl-methylbenzoate (4.7 gm, 0.028mol) and pyridine (5 ml) in methylene chloride (100 ml), a solution ofmethylchloroformate (1.5 eq) was added at 0° C., and the reactionmixture was stirred at room temperature for 1 hr. The reaction mixturewas then poured on ice and extracted with methylene chloride. After theusual workup, the resulting residue was used for the next reaction.

[0101] To a solution of the above compound (0.380 gm, 0.0016 mol.) inCCl₄ (10 ml), NBS was added (0.450 gm, 0.0025 mol.) and the reactionmixture was refluxed until the starting material was completely gone, asevidenced by thin layer chromatography. The succinimide was filtered,and the filtrate was washed with water, dried over Na₂SO₄ andconcentrated under vacuum. The residue was recrystalized from ethylacetate hexane to give 3-methylcarbonate-2-bromomethyl-methylbenzoate(0.415 gm, 85%).

[0102] j) Synthesis of 4-hydroxy-2,3-dihydro-1H-isoindol-1-one (27):

[0103] To a solution of 3-methylcarbonate-2-bromomethyl-methylbenzoate(0.200 gm, 0.65 mmol) in methanol (5 ml) a methanolic ammonia solution(10 ml) was added, and the reaction mixture was stirred at roomtemperature for 2hr. Then the reaction mixture was poured on ice andextracted with ethyl acetate (2×15 ml). After the standard workup theresulting solid was recrystalized in methanol to give compound 27 (0.085gm, 86%).

[0104] 1HNMR (DMSO-D6): 4.20 (s, 2H), 6.94 (d, 1H), 7.09 (d, 1H), 7.26(t, 1H), 8.45 (s, 1H), 9.96 (s, 1 H).

[0105] k) Synthesis of 2,3-dihydro-1H-isoindol-1-one (29):

[0106] To a solution of 4-hydroxy 2,3-dihydro-1H-isoindol-1-one (27)(0.250 gm, 1.67 mmol) in DMF (5 ml) K₂CO₃ and was added, followed byaddition of methyl bromoacetate, and the reaction mixture was stirred atroom temperature for 2hr. Then the reaction mixture was then poured onice and extracted with ethyl acetate (2×15 ml). After the standardworkup, the residue was dissolved in methanol (3 ml), and treated withNaOH (5 ml, 1N) at 0° C. and then stirred for and additional hour. Afterthe standard workup, the solid was recrystalized in methanol to givecompound 29 (0.140 gm, 60%).

[0107] l) Synthesis of isoindolinone (25):

[0108] The suspension of 2,3-dihydro-1H-isoindol-1-one 4 (0.100 gm, 0.61mmol.) in ethyl acetate and aq. NaHCO₃ solution, was addedm-nitrobenzoyl chloride (0.15 mg, 0.81 mmol.) at room temperature. Thereaction mixture was stirred at room temperature for 30 min, and solidseparated was filtered and dried under vacuum to give2,3-dihydro-1H-isoindol-1-one 25 (0.041 gm, 36%).

[0109] m) Synthesis of 2,3-dihydro-1H-isoindol-1-one (32):

[0110] To a solution of 4-hydroxy isoindolinone 27 (0.100 gm, 0.67 mmol)in methylene chloride (5 ml) and triethylamine (0.2 ml), was addedm-nitrobenzoyl chloride (0.125 gm, 0.67 mmol) at 0° C., and the reactionmixture was stirred at room temperature for 5 hr. After the usualworkup, the resulting residue was purified by PTLC plate (solvent was10% methanol-methylene chloride) to give 2,3-dihydro-1H-isoindol-1-one32 (0.025 mg, 28%).

[0111] n) Synthesis of 4-aminomethylene-2,3-dihydro-1H-isoindol-1-one(33):

[0112] To a solution of methyl-2,3-dibromomethylene-benzoate (0.500 gm,0.0015 mol.) in methanol (5 ml), was added methanolic ammonia solution(10 ml), and the reaction mixture was concentrated. The residue obtainedwas dissolved in water (5 ml) and extracted with ethyl acetate. Theaqueous fraction was concentrated under vacuum to give compound 33(0.235 gm, 94%).

[0113] o) Preparation of 4-N-(succinyl)-2,3-dihydro-1H-isoindol-1-one(55):

[0114] A mixture of 4-aminoisoindol-1-one (0.26 gm, 1.8 mmol.) andsuccinic anhydride (0.192 gm, 1.9 mmol.) in anhydrous CHCl₃ (10 ml) wasrefluxed for 2.5 h, and the reaction mixture was left at roomtemperature for 12 h. Then, the solvent was removed under reducedpressure and the residue was triturated with anhydrous ether. A whitecolored solid separated out, which was filtered, washed with anhydrousether and dried under vacuum. Yield 0.36 g (82%).

[0115]¹HNMR(CDCl₃+a few drops of DMSO-D₆): 2.54 (bt, 2H, CH₂), 2.64 (bt,2H, CH₂), 4.30 (s, 2H, CH₂) 7.35-7.55 (m, 2H, Ar—H), 7.70-7.90 (m, 1H,Ar—H), 8.52 (s,1H, exchangeable with D₂O), 9.88 (s,1H, NHCO,exchangeable with D₂O).

[0116] A reaction of variety of anhydrides can be used, such as, forexample, glutaric anhydride (to produce compound 56) or maleic anhydride(to produce compound 35).

[0117] p) A General Procedure for the Preparation of4-N-(diamido)-2,3-dihydro-1H-isoindol-1-one Derivatives (49-51):

[0118] A suspension of the appropriate 4-isoindolin-1-one acid (390μmol) and EDAC-HCl (396 μmol) in dichloromethane (3 ml) was stirred for10 min. Then, diisopropylethylamine (80 μl) and a solution of theappropriate amine (382 μmol) in DMF (0.5 ml) were added. The resultingreaction mixture was stirred at room temperature for 24 h to 72 h.Progress of the reaction was monitored by TLC. After completion, thereaction solvent was removed under reduced pressure and the crudeproduct was purified by prep TLC or by crystallization. A variety ofalkyl or aryl amines were reacted with isoindolin-1-one acid to producethe corresponding derivatives, compounds 49-51.

[0119] q) Preparation of4-N-(tetrahydroisoquinolinosuccinamido)-2,3-dihydro-1H-isoindol-1-one(36):

[0120] A mixture of tetrahydroisoquinoline (2.65 ml, 8 mmol) andsuccinic anhydride (2.06 gm, 20.6 mmol) in anhydrous CHCl₃ (40 ml) wasrefluxed for 2.5 h. Then, the reaction mixture was cooled and dilutedwith dichloromethane (200 ml). The organic layer was washed with citricacid, water and brine and dried over sodium sulfate. The organic layer,on evaporation gave the tetrahydroisoquinolinosuccinic acid derivative.A small amount of the above acid (0.162 gm, 7 mmol) was dissolved in ananhydrous dichloromethane:THF mixture (6 ml, 3/3). To this solutionEDAC-HCl (0.134 gm, 7 mmol), diisopropylethylamine (0.17 ml, 9.8 mmol)and a solution of 4-aminoisoindol-1-one (0.094 gm, 6.4 mmol) in DMF (0.3ml) were added, respectively. The resulting reaction mixture was stirredat room temperature for 120 h. After completion, solvent was removedunder reduced pressure and the residue was triturated with NaHCO₃solution. A solid separated out, which was filtered, washed with water,IN HCl, water and dried under vacuum. Yield 0.175 g (70%).

[0121]¹HNMR(DMSO-D₆): 2.60-3.00 (m, 6H, CH₂), 3.30-3.80 (m, 2H, CH₂),4.30 (s, 2H, CH₂) 4.68 (d, 2H, CH₂), 7.05-7.15 (m, 4H, Ar—H), 7.35-7.45(m, 2H, Ar—H), 7.87 (m, 1H, Ar—H), 8.50 (s,1H, exchangeable with D₂O),9.78 (s,1H, NHCO, exchangeable with D₂O).

[0122] r) Preparation of 4-N-(arginyl)-2,3-dihydro-1H-isoindol-1-one(42):

[0123] To a solution of Boc-Arg (BOC)₂—OH (0.334 gm, 700 μmol) in THF (4ml), EDAC-HCl (0.136 gm, 710 μmol) was added and resulting reactionmixture was stirred at room temperature for 10-15 min. Then,diisopropylethylamine (0.2 ml, 1140 μmol ) and a solution of4-aminoisoindol-l-one (0.1 gm, 668 μmol) in DMF (1 ml) were added,respectively. The resulting reaction mixture was stirred at roomtemperature for 5 days. Then the solvent was removed under reducedpressure and product was extracted with ethyl acetate. The organic layerwas washed with NaHCO₃, brine, dried over sodium sulfate, and evaporatedunder reduced pressure. The residue was treated with 4N HCl-dioxane (2ml) and the resulting mixture was stirred at room temperature for 4 h.After completion, solvent was removed under vacuum, and residue wastriturated with ether. A solid had separated, which was filtered anddried under vacuum. Yield 94 mg (34%).

[0124]¹HNMR(DMSO-d₆+D₂O): 1.40-1.65 (m, 2H, CH₂), 1.80-2.00 (m, 2H,CH₂), 3.05-3.35 (s, 2H, CH₂), 4.08 (t, 1H, CH), 4.40 (s, 2H, CH₂),7.45-7.65 (m, 2H, Ar—H), 7.82 (d, 1H, Ar—H).

[0125] By using this approach, a number of amino acid derivatives of4-aminoisoindolin-1-one were prepared.

[0126] s) Preparation of1-N-{4-(2,3-dihydro-1H-isoindol-1-one)}-3-N-alkyl/aryl-ureas (46 and47):

[0127] To a solution of 4-amino-2,3-dihydro-1H-isoindol-1-one 4 (222μmol) in anhydrous DMF (0.5 ml), a solution of an appropriate alkyl oraryl isocyanate (224 μmol) in dichloromethane was added. The resultingreaction mixture was stirred at room temperature for 48 h. A white solidseparated during the reaction, which was filtered and washed withdichloromethane. The product was dried under vacuum, and characterizedby NMR and mass spectrometry (MS) data.

[0128] 1-N-isoindolinyl-3-alkyl/aryl-urea 47 was also prepared by thisprocedure.

[0129] t) Preparation of 2,3-dihydro-1H-isoindol-1-one (72):

[0130] To a suspension of compound 5 (41.6 gm, 0.18 mol) in CH₃OH (350ml), tert-butyl piperazine carbonate (40 gm, 0.21 mol) was added andstirred further at 50° C. for 48 hr. The TLC of the reaction mixtureshowed completion of the reaction. Then methanol was removed undervacuum. When approximately 100 ml of methanol remained, a white solidprecipitated, which was filtered and washed with cold methanol and driedunder vacuum to give compound 72 (40.2 gm).

[0131] u) Preparation of 2,3-Dihydro-1H-isoindol-1-one (73):

[0132] To a well stirred suspension of 72 (10.1 gm) in CH₃OH (30 ml), anexcess solution of HCl in dioxane (10 ml) was added at 0° C. Thereaction mixture became clear after the addition of the HCl solution,and was left at room temperature for 16 hr. The white precipitate of theHCl salt had separated out, and was diluted with ethyl acetate (10 ml),filtered and washed with cold methanol and dried under vacuum to givecompound 73 (9.2 gm).

[0133] v) Synthesis of isoindolinone (59):

[0134] A suspension of 2,3-dihydro-1H-isoindol-1-one 4 (7 gm, 0.04 mol.)in ethyl acetate and aq. NaHCO₃ solution, was treated withmono-methyloctadioic acid chloride at room temperature. The reactionmixture was stirred at room temperature for 30 min. The solid thatseparated was filtered and dried under vacuum to give2,3-dihydro-1H-isoindol-1-one 85, which was hydrolyzed by KOH (5 eq) inethanol at 0° C. to generate acid 59 (5.775 gm).

[0135] w) Preparation of 2,3-Dihydro-1H-isoindol-1-one (106):

[0136] To a well-stirred suspension of 2′,3′-isopropylidene5′-aminomethyleneadenosine (0.7 gm, 0.002 mol), EDAC (468 mg, 0.002 mol)in DMF (0.5 ml) and CH₂Cl₂ (5 ml), compound 59 (0.7 gm, 0.002 mol) wasadded, followed by addition of diisopropyl ethylamine (0.7 ml) at roomtemperature. The reaction mixture became clear after the addition ofdiisopropyl ethylamine, and urea started precipitating after some time.The reaction was stirred further at room temperature for 24 hrs. Theresidue was purified on a silica gel column using 8% CH₃OH—CH₂Cl₂ togive a protected derivative of compound 106. Water (0.5 ml) was added toa solution of the protected compound (0.9 gm) in TFA (5 ml) and stirredat room temperature for 30 min. The reaction mixture was concentratedunder vacuum (50° C.) to remove the TFA and water. The residue waswashed with ether and ethyl acetate, and recrystalized frommethanol-ethyl acetate to give the TFA salt of 106 (0.575 gm).

[0137] x) Preparation of 2,3-Dihydro-1H-isoindol-1-one (130):

[0138] Compound 73 (32 gm, 0.1 mol) was added to a well stirredsuspension of 2′,3′-isopropylidene adenosine-5′-carboxylic acid (36.5gm, 0.11 mol), DCC (24.5 gm, 0.118 mol), N-hydroxy-succinimide (11.87gm, 0.11 mol) in DMF (200 ml), and was followed by the addition ofdiisopropyl ethylamine (40 ml, 0.22 mol) at room temperature. Thereaction mixture became clear after the addition of diisopropylethylamine, and urea started precipitating after some time. The reactionwas left at room temperature for 3 days. The white precipitate of ureawas filtered and washed thoroughly with DMF. The filtrate wasconcentrated under vacuum to remove diisopropyl ethylamine, and thentreated with acetic acid (2 eq.) to break the DCC complex. Again, thesolid was filtered and discarded. The filtrate was concentrated underhigh vacuum at 60° C., and the residue was purified on the silica gelcolumn using 8% CH₃OH-CH₂Cl₂ to give a of 2′,3′-isopropylidene adenosineprotected derivative of compound 130.

[0139] To a solution of the protected derivative of compound 130 (3.540gm) in TFA (15 ml) was added water (0.4 ml) and the mixture was stirredat room temperature for 1.5 hr. The reaction mixture was concentratedunder vacuum (50° C.) to remove TFA and water. The residue was washedwith ether and ethyl acetate, and recrystalized from methanol-ethylacetate to give the TFA salt of 130 (3.495 gm), which was neutralizedusing a solution of ammonia in methanol (7N) at 0° C., and stirred atroom temperature for 10 min. Next, the reaction was diluted with ethylacetate or ether (10 ml) and the solid was filtered, washed thoroughlywith ethyl acetate or ether and dried under vacuum to yield the freebase of compound 130. See Scheme 1.

[0140] To a suspension of the free base (2.605 gm) in methanol (10 ml),was added a solution of HCl in dioxane (4M, 4 ml) and stirred at roomtemperature for 45 min. Then, it was diluted with ether or ethyl acetate(10 ml) and the solid was filtered, washed thoroughly with ether,recrystalized from CH₃OH, and dried under vacuum to furnish the HCl saltof 130 (2.500 gm).

Example 2

[0141] Effects of substituted 2,3-dihydro-isoindol-1-ones on in vitroinflammation models:

[0142] In in vitro studies, J774 macrophages were exposed toperoxynitrite to induce activation of PARS and related cell injury. Whenthe macrophages were treated with the compounds listed in Table 1,below, the compounds inhibited the activation of PARS and protectedagainst the associated cell necrosis. The results for varioussubstituted isoindolinones are shown in Table 1. TABLE 1

% Cmpd Inhibition No. R (10 μM) 1 H 33 2 F 27 3 NO₂ 12 4 NH₂ 27 5NHCOCH₂Cl 29 6 NHCOCH₂N(CH₃)₂ 66 7 NHCOCH₂N(CH₂CH₃)₂ 74 8NHCOCH₂-4-N-methylpiperazine 66 9 NHCOCH₂CH₂—N-methylpiperazine 59 10NHCOCH₂CH₂CH₂—N-methylpiperazine 35 11 NHCOCH₂NHCH₂CH₃ 66 12NHCOCH₂-piperidine 66 13 NHCOCH₂-1,2,3,4-tetrahydroisoquinoline 80 14NHCOCH₂N(CH₃)CH₂Ph 56 15 NHCOCH₂-(S)-prolinol 83 16 NHCOCH₂-(9-adenine)71 17 NHCOCH₂-9-(6-chloropurine) 75 18NHCOCH₂-9-(6-N,N-dimethylaminpurine) 79 19 NHCOCH₂-benzimidazole 61 20NHCOCH₂-ethylnipecotate 58 21 NHCOCH₂-morpholine 56 22NHCOCH₂-pyrrolidine 51 23NHCOCH₂-1,2,3,4-tetrahydro-6,7-(OCH₃)₂-isoquino- 72 line 24NHCOCH₂-4-indolemethyl ester 78 25 NHCO-m-NO₂-benzoyl 78 26NHCOCH₂CH₂COOC(NH-cyclohexyl)₂ 49 27 OH 60 28 OCH₃ 64 29 OCH₂COOCH₃ 5630 OCH₂COOH 45 31 OCH₂CO-1,2,3,4-tetrahydroisoquinoline 80 32OCO-m-NO₂benzoyl 66 33 CH₂NH₂ 64 34 CH₂NHCO-m-NO₂-benzoyl 45 35NHCOCH═CHCOOH 20 36 NHCO(CH₂)₂—CO-1,2,3,4-tetrahydroisoquinoline 64 37NHCO(CH₂)₃—CO-1,2,3,4-tetrahydroisoquinoline 63 38 NHCONH(CH₂)₃—OH 58 39NHCONH-cyclohexyl 37 40 NHCO(CH₂)₄—NH₂ 55 41NHCOCHNH^(t)Boc-(CH₂)₃—NH—C═NHNtBoc 42 42 NHCOCHNH₂—(CH₂)₃-guanidine 7243 NHCOCHNH^(t)Boc(5-CH₂—N-^(t)Boc-imidazole) 41 44NHCOCHNH^(t)Boc(3-CH₂-indole) 47 45 NHCOCHNH₂(3-CH₂-indole) 60 46NH-prolinyl 46 47 NH-nipecotyl 68 48 NH-3,4,5-(OCH₃)₃-benzoyl 37 49NHCOCH₂CH₂CONH(2-CH₂-benzimidazole) 43 50NHCO(CH₂)₂—CO—NH(CH₂)2-piperidine 42 51NHCO(CH₂)₂—CO—NH(CH₂)₂-2-pyridine 45 52 NHCOCH(NHCOOC₇H₇)(CH₂)₄NHBoc 1453 NHCOCH(NHCOOC₇H₇)(CH₂)₄NH₂ 46 54 NHCOCH₂COOH NT (not tested) 55NHCO(CH₂)₂COOH 25 56 NHCO(CH₂)₃COOH 32 57 NHCO(CH₂)₄COOH NT 58NHCO(CH₂)₅COOH NT 59 NHCO(CH₂)₆COOH NT   60A NHCO(CH2)₇COOH NT   60BNHCO(CH₂)₈COOH NT 61 NHCO(CHOAc)₂COOH NT 62 NHCO(CF₂)₃COOH NT 63NHCONHCH₂COOH NT 64 NHCOCH₂OCH₂COOH NT 65 NHCOCH₂NH₂ NT 66 NHCOCH₂CH₂NH₂NT 67 NHCOCH(CH₃)NH₂ NT 68 NHCOCH₂NHCH₃ NT 69 NHCOCH₂CH₂NHCH₃ NT 70NHCOCH₂CH₂-piperazinetBoc NT 71 NHCOCH₂CH₂NHCH₂CH₃ NT 72NHCOCH₂-N^(t)BOC-Piperazine NT 73 NHCOCH₂-piperazine-HCl 61 74NHCO(CH₂)₇NHCH₃ NT 75 NHCOCH₂CO-piperazine.HCl 43 76 NHCOCH₂CH₂Cl NT 77NHCOCH₂CH₂CH₂Cl NT 78 NHCOC₆H₄—CH═CHCOOH NT 79NHCO(2,5-difluorophenyl)COOH NT 80 NHCOCH₂COOCH₃ NT 81NHCO(CH₂)₂COOCH₂CH₃ 59 82 NHCOCOCOOH NA 83 NHCO(CH₂)₄COOCH₃ 64 84NHCO(CH₂)₅COOCH₃ NT 85 NHCO(CH₂)₆COOCH₃ 73 86 NHCO(p-C₆H₄CH═CHCOOCH₂CH₃)NT 87 NHCO(CH₂)₈COOCH₃ NT 89 NH(2-thiophenoyl) 67 90 NH(2-furoyl) 55 91NH(m-chloromethylenebenzoyl) 40 92 NH(m-N,N-diethylmethylenebenzoyl) 5093 NH(m-NHCOCH₂Cl-benzoyl) 55 94 NH(m-NHCOCH₂N(CH₂CH₃)₂-benzoyl) 45 95NH(m-NHCOCH₂N(CH₃)₂-benzoyl) 50 96NH[m-CH2-(2-hydroxymethylenepyrrolidine)benzoyl] 66 97NH[m-CH2-1,2,3,4-tetrahydroisoquinoline)-benzoyl] 78 98NHCOCH2-isoindoline 38 99 NH(2-carboxybenzoyl) 38 100NHCOCH₂CH(OTBDMS)CH₂COOH NT

Example 3

[0143] Effects of Substituted 2,3-dihydro-isoindol-1-ones on In VivoModels of Reperfusion Injury

[0144] In order to examine the efficacy of the compounds of theinvention in ischemia-reperfusion conditions, the effect of selectedcompounds in a local model of reperfusion injury was examined. Theischemia-reperfusion conditions were induced by ligation and release ofthe superior mesenteric artery in a mouse model. The artery was occludedfor 45 min, followed by reperfusion for 1 h. Following the end of thereperfusion, gut permeability was measured with the fluoresceinisothiocyanate-conjugated dextran (FD4) method in everted gut sacks.See, e.g., Wang et al, J. Surg. Res. 79:39-46, 1998; Wattanasirichaigoonet al., Shock 12: 127-133, 1999; and Tamai et al., Alcohol Clin. Exp.Res. 24: 390-394, 2000. Ischemia-reperfusion increased the permeabilityof the gut from 9±2 to 245±46 ml/min/cm², indicating severe damage ofthe reperfused gut. Treatment with two compounds presented in Table 1(above), where R₁ is H (compound 1) and NHCOCH₂-(S)-prolinol (compound15) (20 mg/kg i.v., injected 10 min prior to the start of reperfusion),reduced the increase in the permeability of the gut to 82±9 and 54±11mlmin/cm², respectively, indicating maintenance of gut function. Theischemia-reperfusion studies in the gut were associated with a <30%mortality, whereas >90% survival was noted in the animals treated withthe compounds. These data indicate that the compounds of the inventionhave therapeutic effects in various systemic and local conditions ofischemia-reperfusion.

Example 4

[0145] Effects of Nucleoside Derivatives on In Vitro InflammationModels:

[0146] The following example illustrates the ability of nucleosidederivatives to inhibit PARS activity. For example, when a nucleosidederivative is conjugated to a compound (such as a substitutedisoindolinone derivative) that may have inherent anti-PARS activity, theconjugate may inhibit the enzyme more strongly that either of the parentmoieties.

[0147] Derivatives with minor modifications to the ribose moiety (e.g.,compounds 137-140 in Table 2) have only modest activity as doesadenosine itself (compound G, Table 3). However, when coupled to cyclicmoieties that have little or modest PARS inhibitory activity alone(e.g., the weaker of the isoindolinone derivatives shown in Table 1),the resultant compound frequently has unexpectedly good PARS inhibitoryactivity.

[0148] For example, conjugates of adenosine derivatives wereinvestigated for PARS inhibition. Compounds 101-113 and 123-130 (Table2) are far more potent than the adenosine derivatives 139 and 140 (Table2) or isoindolinone compounds A and B (Table 3), from which compounds101-113 and 123-130 are derived. It is apparent from compounds 101-130that the nature of L, Z₁ and Z₂, influence the overall potency of theinvention involving coupling of an inhibitor of PARS as group G toadenosine. Compounds 124 and 130 are the most potent derivatives of thisseries.

[0149] Compound 133 (Table 2) is more potent than both the adenosinederivative 139 (Table 2) and the biphenyl compounds F and E (Table 3)from which it was derived.

[0150] Compound 134 (Table 2) is more potent than both the adenosinederivative 139 (Table 2) and the known PARS inhibitor 3-aminobenzamide(E, table 3) from which it was formed.

[0151] Compounds 135 and 136 (Table 2) are more potent than either theadenosine derivative 139 (Table 2) or compound C (Table 3) from whichthey were formed.

Example 5 Effects of Nucleoside Derivatives on In Vivo Inflammation andReperfusion Injury Models:

[0152] In order to examine the efficacy of the compounds of theinvention in ischemia-reperfusion conditions, the effect of selectedcompounds in local models of reperfusion injury was examined.

[0153] Gut ischemia-reperfusion conditions were induced by ligation andrelease of the superior mesenteric artery in a mouse model. The arterywas occluded for 45 min, followed by reperfusion for 1 h. Following theend of the reperfusion, gut permeability was measured with the FD4method in everted gut sacks. Ischemia-reperfusion induced anapproximately 25-fold increase in the permeability of the gut,indicating severe damage of the reperfused gut. Treatment with compounds124 and 130, see Table 2 (below), (10 mg/kg i.v., injected 10 min priorto the start of reperfusion), markedly reduced the increase in thepermeability of the gut to by 72 and 74%, respectively, indicatingmaintenance of the gut function. The ischemia-reperfusion studies in thegut were associated with a <30% mortality without treatment,whereas >80% survival was noted in the animals treated with thecompounds. These data indicate that the compounds of the invention havetherapeutic effects in various systemic and local conditions ofischemia-reperfusion.

[0154] In another set of experiments, the effect of compounds 124 and130 in a rat model of middle cerebral artery occlusion/reperfusion wereexamined. Occlusion lasted for 2 hours, followed by reperfusion for 24hours. Infarct size was quantified with tetrazoliurn staining andsurvival and neurological scores were monitored. The compounds wereadministered at 10 mg/kg i.v. 5 min before the start of reperfusion inone group, and continued at a rate of 10 mg/kg/h infusion during thereperfusion. Vehicle-treated animals (control) developed a >50%mortality over 24 hours, whereas <10% of the animals died in the drugtreated groups. The compounds tested reduced the size of braininfarction by 64±2 and 57±3%, respectively. Vehicle treated animals thatsurvived for 24 h developed severe neurological deficit (3-4 on a scaleof 1-4), whereas in the animals treated with the compounds of theinvention showed minor or no detectable deficit (0-1 on a scale of 1-4).

[0155] In another set of experiments, Male Wistar rats were anesthetizedwith thiopentone sodium (60 mg/kg, i.p.) and were subjected tomyocardial infarction. The chest was opened at the fourth intercostalspace and a 5.0 silk ligature was placed around the left anteriordescending coronary artery for occlusion. Next, one hour of myocardialischemia was performed, followed by 2 h reperfusion. Myocardial infarctsize was quantified by tetrazolium staining. Treatment with compound130, given at 20 mg/kg i.v. 10 min before the start of the reperfusion,reduced the infarct size by 35+7%, as measured at 2 hours ofreperfusion.

[0156] Compounds 124 and 130 were also tested in a mouse model of localinflammation (colitis induced by oral exposure to dextran sulfatesolution, DSS). The compounds were given as oral gavage at 10 mg/kg,twice a day. Vehicle-treated animals (control) developed significantmortality (>60% animals died over 3 weeks), whereas drug treated animalslived longer (<10% mortality at 3 weeks), and exhibited a lesser degreeof intestinal inflammation, as evaluated by gross examination andhistology (severity scores at least 50% reduced by both compounds).Compounds 124 and 130 also protected against endotoxin-induced lethalityin a mouse model (an experimental model of systemic inflammation).

[0157] Taken together, the compounds tested exhibit significantprotective effects in various models of local and systemic inflammationand reperfusion injury, and thus, are likely to exert beneficial effectsin the human equivalent of these diseases. TABLE 2 Cmpd. CompoundInhibition at Inhibition at # Structure 100 μM 10 μM 101

100 64 102

100 100  103

99 88 104

100 67 105

100 66 106

100 98 107

100 62 108

74 34 109

60 29 110

64 29 111

100 75 112

100 73 113

85 55 114

100 56 115

34  0 116

55  0 117

NT NT 118

51.4 31.3 119

58 25 120

100 62 121

81 13.3* 122

NT 25* 123

NT 74* 124

100 at 200 nm 87.1* 125

NT 70* 126

NT 67.1* 127

NT 89* 128

NT 83.2* 129

NT 77.4* 130

100 at 200 nm 97* 131

68 36 132

83 61 133

69  5 134

73 52 135

25 15 136

42 32 137

40 21 138

11.5  0 139

27 NT 140

NT 14.2

[0158] TABLE 3 COM- INHIBITION POUND STRUCTURE at 10 μM A (1)

33 B (4)

27 C

4 D

28 E

35 F

37 G

40

OTHER EMBODIMENTS

[0159] From the above description, one skilled in the art can easilyascertain the essential characteristics of the present invention.Without departing from the spirit and scope thereof, one of ordinaryskill in the art can make various changes and modifications of theinvention to adapt it to various uses and conditions. Other embodimentsare also within the claims.

What is claimed is:
 1. A compound of Formula IV:

or a pharmaceutically acceptable base or acid addition salt, hydrate,ester, solvate, prodrug, metabolite, stereoisomer, or mixtures thereof,wherein: Y is O, OH, S, Se, NH, N-alkyl, N-aryl; R₁₁ is H, OH, aryl,alkyl, or an amino acid side chain; R₁₂ and R₁₃ are, independently,hydrogen, alkyl, aryl, heterocycle, OH, O-alkyl, O-aryl, N-alkyl,N-aryl, or, taken together ═O, ═NH, ═S; R₁₄, R₁₅, R₁₆, and R₁₇ are,independently, hydrogen, halo, alkylhalo, hydroxy, alkoxy, C1-C10straight or branched chain alkyl group, C2-C10 straight or branchedchain alkenyl group, C3-C8 carbocyclic, aryl, alkylamino, amino,carboxy, ester, arylalkyl, nitro groups; or R₁₃, and R₁₄ taken together,form a heterocyclic or a carbocyclic ring; or R₁₄, R₁₅, R₁₆, and R₁₇are, independently, H, or Q—B—C; wherein Q is NHCO, NHCONH, O, CO, OCO₂,OCO, OCONH, NH, CH₂, CH-alkyl, NHCO₂, S, SO₂, CS, SO; B is C1-C10straight or branched chain alkyl, C2-C10 straight or branched chainalkenyl group, C3-C8 carbocyclic, aryl, alkylamino, amino, alkylamido,arylamido, carboxy, ester, anhydride, or arylalkyl group substitutedwith hydrogen, halogen, alkylhalo, hydroxy, nitro, amino, amido,carbamate, or carbonate; C is a substituted carboxy, amido, hydroxy,carbocyclcle or heterocycle, heterocyclic or carbocyclic amine group, ora straight or branched chain alkyl amine, wherein said substitutions areselected from the group consisting of: hydrogen, alkylhalo,alkylhydroxy, C1-C10 straight or branched chain alkyl, C2-C10 straightor branched chain alkenyl group, C2-C10 straight or branched chainalkynyl group, C3-C8 carbocyclic, aryl, benzyl, alkylamino, alkylamido,alkylcarboxy, alkylester, arylalkyl, or cyclic heterocyclic amines, andwherein said heterocyclic amines are selected from the group consistingof piperidine, piperazine, N-alkylated or alkylcarbonylated piperazines,pyrole, imidazole, benzimidazole, tetrazoles, indole, isoquinoline,quinoline, pyrrolidine, aniline, substituted aniline, purine,nucleosides, nucleotides, sugars, hydroxylates alkanes, glycerol, C2 toC10 branched or cyclic or cycloalkenyl amines, and heterocycliccompounds.
 2. The compound of claim 1 wherein Y═O, R11, R12, R13, R15,R16, R17═H and R14 is selected from the group consisting of H, F, NO2,NH2, HCOCH2Cl, NHCOCH2N(CH3)2, NHCOCH2N(CH2CH3)2,NHCOCH2N(CH3)-piperazine, NHCOCH2CH2CH2N(CH3)-piperazine,NHCOCH2NH(CH2CH3), NHCOCH2-piperidine,NHCOCH21,2,3,4-tetrahydroisoquinoline, NHCOCH2N(CH3)CH2Ph,NHCOCH2(S)-prolinol, NHCOCH2-9-adenine, NHCOCH2-9-(6-chloropurine),NHCOCH2-9-(6-N,N-dimethylpurine), NHCOCH2-benzimidazole,NHCOCH2-ethylnipecotate, NHCOCH2-morpholine, NHCOCH2-pyrrolidine,NHCOCH2-1,2,3,4-tetrahydro-6,7-dimethoxyisoquinoline,NHCOCH24-indolemethylester, NHCO-m-NO2-benzoyl,NHCOCH2CH2COOC(NH-cyclohexyl)2, OH, OCH3, OCH2COOCH3, OCH2COOH,OCH2CO-1,2,3,4-tetrahydroisoquinoline, OCO-m-NO2-benzoyl, CH2NH2,CH2NHCO-m-NO2-benzoyl, NHCOCH2CH2CH2COOH, NHCOCH═CHCOOH,NHCOCH2CH2CO-1,2,3,4-tetrahydroisoquinoline,NHCOCH2CH2CH2CO1,2,3,4-tetrahydroisoquinoline, NHCOCH2CH20H,NHCOCH2CH2CH20H, NHCONH-cyclohexyl, NHCO(CH2)4NHtBOC,NHCOCH2CH2CH2CH2NH2, NHCOCHNHtBoc(CH2)3-NH-C═NHN^(t)Boc),NHCOCHNH₂(CH₂)3-guanidine, NHCOCHNH^(t)Boc(5-CH₂-N-tBoc-imidazole),NHCOCHNH^(t)Boc(3-CH₂-indole), NH-prolinyl, NH-nipecotyl,NH-3,4,5-trimethoxybenzoyl, NHCOCH₂CH₂CONH(2-CH₂-benzimidazole),NHCO(CH₂)₂CONH(CH₂)₂-piperidine, NHCO(CH₂)₂CONH(CH₂)₂-piperidine,NHCO(CH₂)₂CONH(CH₂)₂-2-pyridine, NHCOCH(NHCOOC7H7)(CH₂)₄NH^(t)Boc,NHCOCH(NHCOOC₇H₇)(CH₂)₄NH₂, NHCOCH₂COOH, NHCO(CH₂)₂COOH, NHCO(CH₂)₃COOH,NHCO(CH₂)₃COOH, NHCO(CH₂)₄COOH, NHCO(CH₂)₅COOH, NHCO(CH₂)₆COOH,NHCO(CH₂)₇COOH, NHCO(CH₂)₈COOH, NHCO(CHOAc)₂COOH, NHCO(CF₂)₃COOH,NHCONHCH2COOH, NHCOCH20CH2COOH, NHCOCH2NH2, NHCOCH2CH2NH2,NHCOCH(CH3)NH2, NHCOCH2NHCH3, NHCOCH2CH2NHCH3, NHCOCH2tBOC-piperazine,NHCOCH2-piperazine-HCl, NHCO(CH2)7NHCH3, NHCOCH2CO-piperazine-HCl,NHCOCH2CH2Cl, NHCOCH2CH2CH2Cl, NHCOC6H4COCH═CHCOOH,NHCO(2,5-difluorophenyl)COOH, NHCOCH2COOCH3, NHCO(CH2)2COOCH2CH3, NHCOCOCOOCH3, NHCO(CH2)4COOCH3, NHCO(CH2)CH3, NHCO(CH2)6COOCH3,NHCO(p-C6H4-CH═CHCOOCH2CH3), NHCO(CH2)8COOCH3, NH(2-thiophenoyl),NH(2-furoyl), NH(m-chloromethylenebenzoyl),NH(m-N,N-diethylmethylenebenzoyl), NH(m-NHCOCH2Cl-benzoyl),NH(m-NHCOCH2N(CH2CH3)2-benzoyl), NH(m-NHCOCH2N(CH3)2-benzoyl),NH-[m-CH2-(2-hydroxymethylenepyrrolidine)-benzoyl],NH-[m-CH2-(1,2,3,4tetrahydroisoquinoline)-benzoyl], NHCOCH2-isoindoline,NH-2-carboxybenzoyl, and NHCOCH2CH(OTBDMS)-CH2COOH.
 3. The compound ofclaim 1, wherein R₁₃ and R₁₄ together form a heterocyclic or acarbocyclic ring containing from 5 to 10 members.
 4. A method ofinhibiting poly(ADP)-ribose synthase activity in a cell, the methodcomprising contacting said cell with the compound of claim 1 in anamount sufficient to inhibit poly (ADP)-ribose-synthase in said cell. 5.A method of treating or preventing local or systemic inflammation in asubject, the method comprising administering the compound of claim 1 inan amount sufficient to inhibit inflammation in said subject.
 6. Themethod of claim 5, wherein said subject is a human subject.
 7. Themethod of claim 5, wherein said compound is administered systemically.8. The method of claim 5, wherein said compound is administeredtopically.
 9. The method of claim 5, where said local inflammatorycondition is caused by a disorder selected from the group consisting ofan inflammatory disorder of a joint, an inflammatory bowel disease, aninflammatory lung disorder, an inflammatory disease of the centralnervous system, diabetes mellitus, and an inflammatory disease of theeye.
 10. The method of claim 5, wherein said systemic inflammatorycondition is caused by a condition selected from the group consisting ofgram-positive shock, gram-negative shock, hemorrhagic shock,anaphylactic shock, traumatic shock, systemic inflammation, andchemotherapeutic shock.
 11. A method of treating or preventingreperfusion injury in a subject, the method comprising administering thecompound of claim 1 in an amount sufficient to inhibit reperfusioninjury in said subject.
 12. The method of claim 11, wherein saidcompound is administered prophylactically.
 13. The method of claim 11,wherein said compound is administered therapeutically.
 14. The method ofclaim 11, wherein said reperfusion injury is myocardial infarction. 15.The method of claim 11, wherein said reperfusion injury iscardiopulmonary bypass.
 16. The method of claim 11, wherein saidreperfusion injury is stroke.
 17. The method of claim 11, wherein saidsubject is a human subject.
 18. The method of claim 11, wherein saidadministering is topical.
 19. A compound of formula I: A—Z1—L—Z2—G  I ora pharmaceutically accepted prodrug, hydrated salt, or mixtures thereof,wherein: A and G are connected via Z₁ and Z₂ respectively, to a linker,L; A is selected from the group consisting of Formulas II and III;

wherein, X₁ and X₂ are, independently, N or CH, X₃ is CR₇═CR₈,CHR₇—CHR₈, CR₇═N, N═CR₇, N═N, NR₇—O, CHR₇—O or CHR₇—S, where R₇ and R₈are, independently, H, alkyl, amino, substituted amino, hydroxy, oralkoxy; R₁ and R₂ are independently H, alkyl, NH2, OH, SH, Cl, NHR₉,N═R₉, N═NR₉, or amide, where R₉ is alkyl, aryl, arylalkyl,alkyl-heterocycle; R₃ and R₄ are independently H, hydroxy, C1-5 alkyl,amino, or halo; R₅ and R₆ are independently alkyl, acetyl, other acyl,or R₅ and R₆ taken together form a 5-or 6-membered optionallysubstituted heterocycle; Z₁ is —CH₂O—, —CH₂NR₁₀—, —CH₂NR₁₀C(O)—,—CONR₁₀—, —CO₂—, —CH₂NHCONH—, —CH₂—, —CH₂NHCSNH—, —CO—, —CH₂CO₂—,—NHCO₂, S, SO₂, CH₂S, or SO; Z₂ is; —NR₁₀CO—, —C(O)NR₁₀—, —NHCONH—,—OC(O)—, —C(O)O—, NHCS, —CSNH—, NHCSNH, O, CO, OCO, OCONH, NH, CH₂,CH-alkyl, NHCO₂, S, SO₂, CS, or SO; L is H, O, S, C1-15 alkylene chainoptionally substituted in one or more positions, or a 5, 6 or 7 memberedcarbocycle or heterocycle optionally substituted in one or morepositions, provided that L is not H when Z₁ is O, and when L═H, Z₂ and Gare absent, and G is H, OH, SH, NH₂, CO₂H, unsubstituted or substitutedalkyl, aryl, alkylaryl, carbocyclic, heterocyclic, bicyclic, bicyclicheterocycle, biphenyl or heterocyclic amines such as piperidine,piperazine, pyrole, imidazole, benzimidazole, tetrazoles, indole,isoquinoline, quinoline, or pyrrolidine.
 20. The compound of claim 19,wherein group A is a pyrimidine or purine nucleoside or derivative. 21.The compound of claim 19, wherein group G is an inhibitor of PARS. 22.The compound of claim 19, wherein Z₁ is C(O)N(R₁₀)—, and Z₂ is —C(O) NH.23. The compound of claim 19, wherein group L is a C1-C10 carbon chainor a heterocyle.
 24. The compound of claim 19, wherein G is an inhibitorof PARS and A is a pyrimidine or purine nucleoside or derivative. 25.The compound of claim 19, wherein G is an inhibitor of PARS and A isadenosine, a derivative of adenosine or an analog of adenosine.
 26. Thecompound of claim 19, wherein G is an isoindolinone and A is adenosineattached to Z₁ at the 5′ position.
 27. The compound of claim 26, whereinZ₁ is C(O)N(CH₃)—and Z₂ is —C(O) NH.
 28. The compound of claim 27,wherein L is CH₂CH₂.
 29. The compound of claim 26, wherein Z₁ is —CO—,Z₂ is —NHCO— and L is -(5′-methylene)-N-piperazyl.